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Based on the developing trend of laboratory medicine and the internal rules of discipline construction,series of research and reform practices in the curriculum system were put forward,including practice teaching,innovation education and instructional technology for laboratory medicine.An innovation talents training mode for laboratory medicine is gradually formed and innovative education is pushed on systematically,effectively and abidingly.The new mode has made periodical results and good prospects in cultivating students with innovative quality and ability.
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<p><b>OBJECTIVE</b>To develop a method for extracting cytoskeletons and cytoskeleton-associated proteins for proteomic analysis.</p><p><b>METHODS</b>A subcellular sequential proteome extraction method was exploited. The extraction procedure was optimized and controlled according to observed cell morphology changes and one- and two-dimensional electrophoresis images. The extraction efficiency and selectivity were evaluated by Western blotting and mass spectrometry.</p><p><b>RESULTS</b>Four extracted fractions clearly displayed distinct patterns. Western blotting detected the fraction-marker proteins FAK, integrin-β1, histone H1 and cytokeratin 19 only in their expected fractions. About 90% of the protein spots in the cytoskeleton fraction were identified by mass spectrometry as cytoskeleton and/or its associated proteins.</p><p><b>CONCLUSION</b>The subcellular proteome sequential fractionation method facilitates the detection of proteins of low abundance and shows a high reproducibility and selectivity, and thus can serve as an ideal pre-fractionation method prior to two-dimensional electrophoresis.</p>
Subject(s)
Humans , Cytoskeleton , Chemistry , Electrophoresis, Gel, Two-Dimensional , Methods , Proteome , Proteomics , Methods , Subcellular FractionsABSTRACT
Cultivation of young teachers is an important part of teaching staff training in colleges and universities. To strengthen teaching ability of young teachers in discipline of medicine laboratory is related to the quality of laboratory medicine personnel training.The paper will be based on some problems of young teacher teaching and discuss some measures of young teachers' teaching ability improvement.
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ObjectiveTo explore the effect and mechanism of Toll-like receptor 4(TLR4) ligand LPS-mediated inhibition hepatitis B virus (HBV) replication in Bewo cells.MethodsFirst of all,2 μg 1.3-fold HBV recombinant vector pcDNA3.1 ( + )-HBV1.3 were transfected into Bewo cells,after 12 h,the cells were treated with LPS for 3 d.To observe the kinetics of IFN-β and TNF-α expression in Bewo cells,the Bewo cells were exposed to TLR4 ligand LPS.And the effect of pyrrolidine dithiocarbamate ( PDTC),an inhibitor of NF-κB,on LPS-induced cytokines was also observed.The HBsAg,HBeAg and HBV DNA level in the culture supernatant were detected by Microparticle Enzyme Immunoassay (MEIA) and fluorescence quantitative PCR,respectively,and the expression of IFN-β,TNF-α,TRIF and MyD88 was detected by ELISA and RT-PCR,respectively.ResultsCompared with control group,LPS could significantly suppress HBV replication in Bewo cells ( P ＜0.01 ),and it could induce the production of TNFα in Bewo cells ( P ＜ 0.05 ),in time-and dose-dependent manners.PDTC strongly inhibited LPS and induced TNF-α production,but had no much effect on IFN-β in Bewo cells ( P ＜ 0.001 ).Compared with control group,the mRNA levels of MyD88 were significantly induced by LPS in the Bewo cells transfected with this recombinant vector( P ＜ 0.001 ).ConclusionsTLR4 ligand LPS could significantly suppress HBV replication by inducing TNF-α production in Bewo cells mainly via the MyD88/ NF-κB signal pathway.
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Objective To explore the changes and clinical significance of plasma tissue factor pathway (TFP) levels during the onset of acute ischemic cardiac and cerebral vessel disease. Methods The study population consisted of 69 patients with AMI and 71 with AICS as well as 50 age-matched healthy volunteers. Blood samples were obtained during the onset period of AMI and AICS. Tissue factor (TF) and tissue factor pathway inhibitor (TFPI) activity in plasma were assayed with the chromogenic assay, Plasma TF and TFPI antigen were measured with ELISA, Plasma FⅦ coagulation activity (FⅦ:C) was developed in the one- stage system. Plasma FⅦa was determined by soluble TF assay, plasma prothrombin (FII) and fibrinogen (Fbg) were measured by one-stage amidolytic assay and thrombin time (TT) respectively. Results In the AMI group, compared with the control group, plasma TF activities and antigens, TFPI activities and antigens as well as FⅦa were significantly higher(all P0.05), plasma FII and Fbg were also remarkably increased (both P0.05). Conclusion The initiation of TF pathway would be associated with the onset of AMI and AIS, but the changes of TF, TFPI, FⅦ:C and Fbg in AMI are different from those in AICS, endothelial dysfunction and the blood is in hypercoagulable state.